Methylthioadenosine deaminase in an alternative quorum sensing pathway in Pseudomonas aeruginosa

Rong Guan, Meng Chiao Ho, Richard F G Fröhlich, Peter C. Tyler, Steven C. Almo, Vern L. Schramm

Research output: Contribution to journalArticle

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Abstract

Pseudomonas aeruginosa possesses an unusual pathway for 5′-methylthioadenosine (MTA) metabolism involving deamination to 5′-methylthioinosine (MTI) followed by N-ribosyl phosphorolysis to hypoxanthine and 5-methylthio-α-d-ribose 1-phosphate. The specific MTI phosphorylase of P. aeruginosa has been reported [Guan, R., Ho, M. C., Almo, S. C., and Schramm, V. L. (2011) Biochemistry 50, 1247-1254], and here we characterize MTA deaminase from P. aeruginosa (PaMTADA). Genomic analysis indicated the PA3170 locus to be a candidate for MTA deaminase (MTADA). Protein encoded by PA3170 was expressed and shown to deaminate MTA with 40-fold greater catalytic efficiency for MTA than for adenosine. The kcat/K m value of 1.6 × 107 M-1 s-1 for MTA is the highest catalytic efficiency known for an MTA deaminase. 5′-Methylthiocoformycin (MTCF) is a 4.8 pM transition state analogue for PaMTADA but causes no significant inhibition of human adenosine deaminase or MTA phosphorylase. MTCF is permeable to P. aeruginosa and exhibits an IC 50 of 3 nM on cellular PaMTADA activity. PaMTADA is the only activity in P. aeruginosa extracts to act on MTA. MTA and 5-methylthio-α-d-ribose are involved in quorum sensing pathways; thus, PaMTADA is a potential target for quorum sensing. The crystal structure of PaMTADA in complex with MTCF shows the transition state mimic 8(R)-hydroxyl group in contact with a catalytic site Zn2+, the 5′-methylthio group in a hydrophobic pocket, and the transition state mimic of the diazepine ring in contact with a catalytic site Glu.

Original languageEnglish (US)
Pages (from-to)9094-9103
Number of pages10
JournalBiochemistry
Volume51
Issue number45
DOIs
StatePublished - Nov 13 2012

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Quorum Sensing
Pseudomonas aeruginosa
Methylthioinosine
Catalytic Domain
5'-methylthioadenosine
Phosphorylases
Deamination
Biochemistry
Hypoxanthine
Ribose
Metabolism
Hydroxyl Radical
Adenosine
Crystal structure

ASJC Scopus subject areas

  • Biochemistry

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Methylthioadenosine deaminase in an alternative quorum sensing pathway in Pseudomonas aeruginosa. / Guan, Rong; Ho, Meng Chiao; Fröhlich, Richard F G; Tyler, Peter C.; Almo, Steven C.; Schramm, Vern L.

In: Biochemistry, Vol. 51, No. 45, 13.11.2012, p. 9094-9103.

Research output: Contribution to journalArticle

Guan, Rong ; Ho, Meng Chiao ; Fröhlich, Richard F G ; Tyler, Peter C. ; Almo, Steven C. ; Schramm, Vern L. / Methylthioadenosine deaminase in an alternative quorum sensing pathway in Pseudomonas aeruginosa. In: Biochemistry. 2012 ; Vol. 51, No. 45. pp. 9094-9103.
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abstract = "Pseudomonas aeruginosa possesses an unusual pathway for 5′-methylthioadenosine (MTA) metabolism involving deamination to 5′-methylthioinosine (MTI) followed by N-ribosyl phosphorolysis to hypoxanthine and 5-methylthio-α-d-ribose 1-phosphate. The specific MTI phosphorylase of P. aeruginosa has been reported [Guan, R., Ho, M. C., Almo, S. C., and Schramm, V. L. (2011) Biochemistry 50, 1247-1254], and here we characterize MTA deaminase from P. aeruginosa (PaMTADA). Genomic analysis indicated the PA3170 locus to be a candidate for MTA deaminase (MTADA). Protein encoded by PA3170 was expressed and shown to deaminate MTA with 40-fold greater catalytic efficiency for MTA than for adenosine. The kcat/K m value of 1.6 × 107 M-1 s-1 for MTA is the highest catalytic efficiency known for an MTA deaminase. 5′-Methylthiocoformycin (MTCF) is a 4.8 pM transition state analogue for PaMTADA but causes no significant inhibition of human adenosine deaminase or MTA phosphorylase. MTCF is permeable to P. aeruginosa and exhibits an IC 50 of 3 nM on cellular PaMTADA activity. PaMTADA is the only activity in P. aeruginosa extracts to act on MTA. MTA and 5-methylthio-α-d-ribose are involved in quorum sensing pathways; thus, PaMTADA is a potential target for quorum sensing. The crystal structure of PaMTADA in complex with MTCF shows the transition state mimic 8(R)-hydroxyl group in contact with a catalytic site Zn2+, the 5′-methylthio group in a hydrophobic pocket, and the transition state mimic of the diazepine ring in contact with a catalytic site Glu.",
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